Therapeutic treatment

ABSTRACT

A combination comprising candesartan and rosuvastatin for the prevention or treatment of atherosclerosis and for the prevention of cardiovascular events is described.

The present invention relates to a combination comprising candesartanand rosuvastatin.

The present invention further relates to pharmaceutical compositionscomprising the combination mentioned hereinbefore. The present inventionfurther relates to the use of a combination mentioned hereinbefore inthe prevention or treatment of atherosclerosis.

Atherosclerosis is a condition mediated by complex pathologicalprocesses which result in irregularly distributed lipid deposits in thearteries and is a major contributory factor to coronary heart disease. Areduction in atherosclerosis is therefore a major target for reducingthe number of cardiovascular events for example, myocardial infarction,worsening of angina, cardiac arrest, stroke, congestive heart failureand cardiovascular death.

Dyslipidemia, particularly increased plasma level of low-densitylipoprotein (LDL) is one of the major risk factors in atherosclerosis.Clinical studies have demonstrated that reducing plasma LDL level with3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors,commonly known as statins, results in a lower risk of cardiovascularevents.

Activation of the renin-angiotensin system (RAS) may be consideredanother important risk factor in atherosclerosis. Activation of RAS withthe formation of angiotensin (II) (A (II)) and the activation of A (II)receptors have been implicated in atherogenesis, plaque rupture,myocardial ischemic dysfunction and congestive heart failure (Singh andMehta, Arch Intern Med, 2003, vol 163, 1296-1304).

International Patent Application WO 95/26188 discloses treatment ofatherosclerosis with A(II) receptor blockers, optionally in combinationwith HMGCoA reductase inhibitors. International Patent Application WO01/76573 discloses the use of a combination of at least two of an A(II)antagonist, an ACE (angiotensin converting enzyme) inhibitor and anHMGCoA reductase inhibitor for the prevention or delay of progression ina list of conditions, amongst which is atherosclerosis.

We have surprisingly found that the combination of the A(II) antagonistcandesartan and the HMG CoA reductase inhibitor rosuvastatin has asynergistic effect in the reduction of atherosclerosis. This synergisticeffect appears to arise from synergistic inhibition of expression of anumber of inflammatory mediators involved in the RAS (for example CD40,metalloproteinases (MMPs)) and/or inhibition of the expression of thereceptor LOX-1 (which is a receptor for oxidised LDL on endothelialcells). The synergistic effect provides strong evidence for cross-talkbetween the RAS and dyslipidemia in atherogenesis.

It will be appreciated that the activity of MMPs may be regulatedin-vivo by their tissue inhibitors (TIMPs). We have also shown that theexpression of TIMP-1 and TEMP-2 is up-regulated by high-cholesteroldiet, and markedly attenuated by the combination of candesartan androsuvastatin. These data lend credence to the concept that the balancebetween MMPs and TIMPs is altered by high-cholesterol diet, and thatthis imbalance can be “normalized” by the combination of an A(II)antagonist and a lipid-lowering agent.

In one aspect of the invention is provided a combination comprisingcandesartan, or a pharmaceutically acceptable salt thereof, androsuvastatin, or a pharmaceutically acceptable salt thereof, for theprevention or treatment of atherosclerosis.

In one aspect of the invention is provided a combination comprisingcandesartan, or a pharmaceutically acceptable salt thereof, androsuvastatin, or a pharmaceutically acceptable salt thereof, for theprevention of cardiovascular events.

Such a combination may also be useful in the treatment or prevention ofother diseases associated with these mediators, for example ininflammatory diseases or conditions, such as ischemia-reperfusion injury(to the heart, brain, kidneys, lungs and liver), radiation-inducedinjury, burn injury and peripheral vascular disease,

Candesartan may suitably be in the form of candesartan, or in thepro-drug form candesartan cilexetil. These forms may be formulated witha further agent such as a diuretic such as hydrochlorothiazide (forexample, as marketed as Atacand Plus™).

Where herein candesartan is referred to, this includes both candesartanand candesartan cilexetil.

Preferably the calcium salt of rosuvastatin, which may be referred to asrosuvastatin calcium, is used in the various aspects of the presentinvention.

In general, pharmaceutically-acceptable salts include acid additionsalts such as methanesulfonate, tosylate, α-glycerophosphate, fumarate,hydrochloride, citrate, maleate, tartrate and (less preferably)hydrobromide. Pharmaceutically-acceptable salts in general also includesalts formed with phosphoric and sulfuric acid.Pharmaceutically-acceptable salts generally include base salts such asan alkali metal salt for example sodium, an alkaline earth metal saltfor example calcium or magnesium, an organic amine salt for exampletriethylamine, morpholine, N-methylpiperidine, N-ethylpiperidine,procaine, dibenzylamine, N,N-dibenzylethylamine,tris-(2-hydroxyethyl)amine, tris(hydroxymethyl)methylammonium, N-methyld-glucamine and amino acids such as lysine. There may be more than onecation or anion depending on the number of charged functions and thevalency of the cations or anions.

Herein, where the term “combination” is used it is to be understood thatthis refers to simultaneous, separate or sequential administration. Inone aspect of the invention “combination” refers to simultaneousadministration. In another aspect of the invention “combination” refersto separate administration. In a further aspect of the invention“combination” refers to sequential administration. Where theadministration is sequential or separate, the delay in administering thesecond component should be such that both agents are present in the bodyso as to produce the synergistic effect of the combination.

In a further aspect of the invention is provided a pharmaceuticalcomposition which comprises candesartan, or a pharmaceuticallyacceptable salt thereof, and rosuvastatin, or a pharmaceuticallyacceptable salt thereof, in association with a pharmaceuticallyacceptable diluent or carrier for use in the prevention or treatment ofatherosclerosis.

In a further aspect of the invention is provided a pharmaceuticalcomposition which comprises candesartan, or a pharmaceuticallyacceptable salt thereof, and rosuvastatin, or a pharmaceuticallyacceptable salt thereof, in association with a pharmaceuticallyacceptable diluent or carrier for use in the prevention or reduction ofrisk of cardiovascular events.

The compositions described herein may be in a form suitable for oraladministration, for example as a tablet or capsule, for parenteralinjection (including intravenous, subcutaneous, intramuscular,intravascular or infusion) for example as a sterile solution, suspensionor emulsion, for topical administration for example as an ointment orcream, for rectal administration for example as a suppository or theroute of administration may be by direct injection into the tumour or byregional delivery or by local delivery. In other embodiments of thepresent invention the compounds of the combination treatment may bedelivered endoscopically, intratracheally, intralesionally,percutaneously, intravenously, subcutaneously, intraperitoneally orintratumourally. In general the compositions described herein may beprepared in a conventional manner using conventional excipients orcarriers that are well known in the art.

Suitable pharmaceutically-acceptable excipients or carriers for a tabletformulation include, for example, inert excipients such as lactose,sodium carbonate, calcium phosphate or calcium carbonate, granulatingand disintegrating agents such as corn starch or alginic acid; bindingagents such as gelatin or starch; lubricating agents such as magnesiumstearate, stearic acid or talc; preservative agents such as ethyl orpropyl 4-hydroxybenzoate, and anti-oxidants, such as ascorbic acid.Tablet formulations may be uncoated or coated either to modify theirdisintegration and the subsequent absorption of the active ingredientwithin the gastrointestinal tract, or to improve their stability and/orappearance, in either case using conventional coating agents andprocedures well known in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid excipient, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin or olive oil.

Candesartan is commercially available as ‘Atacand™’ and ‘Atacand Plus™’Rosuvastatin calcium is commercially available as ‘Crestor™’. Suitableformulations for the present invention include those which arecommercially available.

Suitable dosages of each component of the combination are those of themarketed commercial products. Alternatively, the synergy between thecomponents may allow a lower dosage of one or both components to beused. For example, a dose of 4 mg, 8 mg, 16 mg, 32 mg, or up to 160 mgof candesartan in combination with a dose of 80 mg, 40 mg, 20 mg, 10 mg,5 mg or 2.5 mg of rosuvastatin may be used. It will be understood thatany one of the doses of candesartan may be combined with any suitabledose of rosuvastatin.

In one aspect, 80 mg of rosuvastatin is used. In another aspect, 40 mgof rosuvastatin is used. In a further aspect, 20 mg of rosuvastatin isused. In a further aspect, 10 mg of rosuvastatin is used. In a furtheraspect, 5 mg of rosuvastatin is used. In a further aspect, 2.5 mg ofrosuvastatin is used.

In one aspect, between 32 and 160 mg, such as about 64 to 128 mg, forexample 64 to 112 mg, such as about 64-96 mg of candesartan is used.Conveniently, about 72 mg of candesartan is used. In another aspect, 32mg of candesartan is used. In a further aspect, 16 mg of candesartan isused. In a further aspect, 8 mg of candesartan is used. In a furtheraspect, 4 mg of candesartan is used.

It will be appreciated that the pharmaceutical composition according tothe present invention includes a composition comprising candesartan or apharmaceutically acceptable salt thereof and rosuvastatin or apharmaceutically acceptable salt thereof and apharmaceutically-acceptable excipient or carrier. Such a composition,for example in a single oral formulation conveniently provides thetherapeutic combination product of the invention for simultaneousadministration in the prevention or treatment of atherosclerosis.

Preferably the two components of the combination are both administeredorally.

Preferably the two components of the combination are administered as asingle oral formulation.

Preferably the combination is formulated for once-a-day dosing.

Conveniently, the combination is formulated as a single tablet orcapsule.

The dosages and schedules described hereinbefore may be varied accordingto the particular disease state and the overall condition of thepatient. For example, it may be necessary or desirable to reduce theabove-mentioned doses of the components of the combination treatment inorder to reduce toxicity. Dosages and schedules may also vary if, inaddition to a combination treatment of the present invention, one ormore additional chemotherapeutic agents are used. Scheduling can bedetermined by the practitioner who is treating any particular patientusing his professional skill and knowledge.

A pharmaceutical composition according to the present invention alsoincludes separate compositions comprising a first composition comprisingcandesartan or a pharmaceutically acceptable salt thereof and apharmaceutically-acceptable excipient or carrier, and a secondcomposition comprising rosuvastatin or a pharmaceutically acceptablesalt thereof and a pharmaceutically-acceptable excipient or carrier.Such a composition conveniently provides the therapeutic combination ofthe invention for sequential or separate administration in thesynergistic prevention or treatment of atherosclerosis but the separatecompositions may also be administered simultaneously.

In another aspect of the invention there is provided a combinationcomprising candesartan, or a pharmaceutically acceptable salt thereof,and rosuvastatin, or a pharmaceutically acceptable salt thereof, for useas a medicament for the prevention or treatment of atherosclerosis.

In another aspect of the invention there is provided a combinationcomprising candesartan, or a pharmaceutically acceptable salt thereof,and rosuvastatin, or a pharmaceutically acceptable salt thereof, for useas a medicament for the prevention of cardiovascular events.

In another aspect of the invention there is provided a combinationcomprising candesartan, or a pharmaceutically acceptable salt thereof,and rosuvastatin, or a pharmaceutically acceptable salt thereof, for usein the manufacture of a medicament.

In another aspect of the invention there is provided a combinationcomprising candesartan, or a pharmaceutically acceptable salt thereof,and rosuvastatin, or a pharmaceutically acceptable salt thereof, for usein the manufacture of a medicament for the prevention or treatment ofatherosclerosis.

In another aspect of the invention there is provided a combinationcomprising candesartan, or a pharmaceutically acceptable salt thereof,and rosuvastatin, or a pharmaceutically acceptable salt thereof, for usein the manufacture of a medicament for the prevention of cardiovascularevents.

In a further aspect of the invention there is provided a method ofpreventing or treating atherosclerosis in a warm-blooded animal, such asman, which comprises administering a combination of candesartan, or apharmaceutically acceptable salt thereof, and rosuvastatin, or apharmaceutically acceptable salt thereof.

In a further aspect of the invention there is provided a method ofpreventing cardiovascular events in a warm-blooded animal, such as man,which comprises administering a combination of candesartan, or apharmaceutically acceptable salt thereof, and rosuvastatin, or apharmaceutically acceptable salt thereof.

According to a further aspect of the present invention there is provideda kit comprising a combination of candesartan or a pharmaceuticallyacceptable salt thereof, and rosuvastatin; or a pharmaceuticallyacceptable salt thereof, optionally with instructions for use in theprevention or treatment of atherosclerosis.

According to a further aspect of the present invention there is provideda kit comprising:

a) candesartan in a first unit dosage form;b) rosuvastatin in a second unit dosage form; andc) container means for containing said first and second dosage forms;and optionallyd) with instructions for use in the prevention or treatment ofatherosclerosis.

According to another aspect of the present invention there is provided amethod of inhibiting expression of CD40 and/or metalloproteinases (MMPs)by administering a combination of candesartan, or a pharmaceuticallyacceptable salt thereof and rosuvastatin, or a pharmaceuticallyacceptable salt thereof.

Particular metalloproteinases are MMP-1, MMP-2 and MMP-9. According toanother aspect of the present invention there is provided a method oftreating atherosclerotic patients by inhibition of expression of CD40and/or metalloproteinases (MMPs) by administering an amount of acombination of candesartan, or a pharmaceutically acceptable saltthereof and rosuvastatin, or a pharmaceutically acceptable salt thereofsuitable for inhibition of expression of CD40 and/or metalloproteinases(MMPs).

According to a further aspect of the invention, there is provided amethod for normalizing the balance between MMPs and TIMPS byadministration of an amount of a combination of candesartan, or apharmaceutically acceptable salt thereof and rosuvastatin, or apharmaceutically acceptable salt thereof.

According to another aspect of the present invention there is provided amethod of inhibiting expression of LOX-1 by administering a combinationof candesartan, or a pharmaceutically acceptable salt thereof androsuvastatin, or a pharmaceutically acceptable salt thereof.

According to another aspect of the present invention there is provided amethod of treating atherosclerotic patients by inhibition of expressionof LOX-1 by administering an amount of a combination of candesartan, ora pharmaceutically acceptable salt thereof and rosuvastatin, or apharmaceutically acceptable salt thereof suitable for inhibition ofexpression of LOX-1.

Materials and Methods Animal Model

Five pairs of C57BL/6J mice and three pairs of homozygous apo-E knockoutmice (on C57BL/6J background) were obtained from Jackson Laboratories(Bar Harbor, Me.). They were bred by brother-sister mating and housed ina room lit from 6:00 AM to 6:00 PM and kept at 21° C. The C57BL/6J mice(n=10) were continued on regular diet for the entire study period. Theapo-E knockout mice were divided into four groups. Group 1 (n=10)animals were given high-cholesterol diet (1% cholesterol) alone for 12weeks since the age of 6 weeks; Group 2 (n=10) animals were givenhigh-cholesterol diet with candesartan (1 mg/kg/d) for 12 weeks sincethe age of 6 weeks; Group 3 (n=10) animals were given high-cholesteroldiet with the rosuvastatin (1 mg/kg/d) for 12 weeks since the age of 6weeks; Group 4 (n=10) animals were given high-cholesterol diet withcandesartan (1 mg/kg/d) and rosuvastatin (1 mg/kg/d) for 12 weeks sincethe age of 6 weeks.

At the end of 12-week-treatment, the mice were sacrificed and subject tostudies described below. All experimental procedures were performed inaccordance with protocols approved by the Institutional Animal Care andUsage Committee of University of Arkansas for Medical Sciences.

Quantitative Analysis of Atherosclerotic Plaques

At the end of 12-week-treatment, 5 mice from each group were euthanizedand the aortas were separated from surrounding tissues. After removal ofthe adventitial fat tissue, the aortas were opened longitudinally fromthe aorta arch to the iliac bifurcation, and fixed in 10% formalin for24 hours. Then the aortas were rinsed in 70% alcohol briefly, stainedwith Sudan IV solution for 15 minutes, differentiated in 80% alcohol for20 minutes and washed in running water for 1 hour (Russell L. Techniquesfor studying atherosclerotic lesion, Lab Invest. 1958; 7:42-47). Theaortas were mounted and their pictures were taken with a cameraconnected to a dissection microscope. The images were analyzed by software (Image Pro Plus, Media Cybernetics).

RNA Preparation and Analysis by RT-PCR

At the end of 12-week-treatment, 5 mice from each group were euthanizedand the aortas (from aorta arch to iliac bifurcation) were separatedfrom surrounding tissues and stored on dry ice. Each aorta was cut intofour segments, two of which were used to extract total RNA with thesingle-step acid-guanidinium thiocyanate-phenol-chloroform method asdescribed earlier (27). One microgram of total RNA was reversetranscripted into cDNA with oligo-dT (Promega, Madison, Wis., U.S.A.)and Maloney murine leukemia virus (M-MLV) reverse transcription(Promega) at 42° C. for 1 hour. Two microliters of reverse transcription(RT) material was amplified with Taq DNA polymerase (Promega) and aprimer pair specific to mouse LOX-1, CD40 or MMPs (MMP-1, -2, -9). Formouse LOX-1, forward primer: 5′-TTACTCTCCATGGTGGTGCC-3′, reverse primer:5′-AGCTTCTTCTGCTTGTTGCC-3′ were used. 30 cycles of polymerase chainreaction (PCR) were performed at 94° C. for 40 seconds (denaturation),55° C. for 1 minute (annealing), and 72° C. for 1 minute (extension).The size of polymerase chain reaction (PCR) product was 193 base pairs.For mouse CD40, forward primer 5′-GTTTAAAGTCCCGGATGCGA-3′ and reverseprimer 5′-CTCAAGGCTATGCTGTCTGT-3′ were used. 35 cycles of polymerasechain reaction (PCR) were performed at 94° C. for 1 minute(denaturation), 55° C. for 1 minute (annealing), and 72° C. for 1 minute(extension). The size of PCR product was 408 base pairs. For mouseMMP-1, forward primer 5′-GGACTCTCCCATTCTTAATGA T-3′ and reverse primer5′-CCTCTTTCTGGATAACATCATCA AC-3′ were used. For mouse MMP-2, forwardprimer 5′-ATCAAGGGGATCCAGGAGC-3′ and reverse primer 5′-GCAGCGATGAAGATGATAG-3′ were used. For mouse MMP-9, forward primer5′-AGTTTGGTGTCGCGGAGCAC-3′ and reverse primer 5′-TACATGAGCGCTTCCGGCAC-3′were used. For all MMPs, 35 cycles of PCR were performed at 94° C. for 1minute (denaturation), 58° C. for 1 minute (annealing), and 75° C. for 1minute (extension). The sizes of PCR product were 627, 718 and 753 basepairs, respectively. A primer pair specific to mouse β-actin was used ashousekeeping gene (forward primer: 5′-TTCTACAATGAGCTGCGTTG-3′, reverseprimer: 5′-CACTGTGTTGGCATAGAGGTC-3′), 30 cycles were used at 94° C. for30 seconds (denaturation), 55° C. for 1 minute (annealing), and 72° C.for 1 minute (extension). PCR product was 560 base pairs. The reversetranscription PCR(RT-PCR)-amplified sample was visualized on 1.5%agarose gel using ethidium bromide.

Protein Preparation and Analysis by Western Blot

Each mouse aorta was cut into four segments. Two of them were used toextract RNA, and the remaining two were used to extract protein asdescribed previously (14). In brief, the aortic tissues were homogenizedand lysed in lysis buffer, then centrifuged at 4000 rpm for 10 minutesat 4° C. The lysate proteins from aortas (20 μg/lane) were separated by10% SDS-PAGE, and transferred to nitrocellulose membranes. Afterincubation in blocking solution (5% non-fat milk, Sigma), membranes wereincubated with 1:750 dilution monoclonal antibody to mouse LOX-1, 1:500dilution polyclonal antibody to mouse CD40 (Santa Cruz), 1 μg/mldilution monoclonal antibody to mouse MMP-1 (Oncogene), 1 μg/ml dilutionmonoclonal antibody to mouse MMP-2 (Oncogene), 1 μg/ml dilutionmonoclonal antibody to mouse MMP-9 (Oncogene), 1:500 dilution polyclonalantibody to mouse TIMP-1 (Santa Cruz), 1:500 dilution polyclonalantibody to mouse TIMP-2 (Santa Cruz), or 1:5000 dilution monoclonalantibody to mouse β-actin (Sigma) for overnight at 4° C. Membranes werewashed and then incubated with 1:5000 dilution specific secondaryantibody (Amersham Life Science) for 2 hours at room temperature, andthe membranes were washed and detected with the ECL system (AmershamLife Science). The relative intensities of protein bands were analyzedby Scan-gel-it software (Li D Y, Zhang Y C, Sawamura T, Mehta J L. CircRes. 1999; 84:1043-1049).

Data Analysis

All data represent mean of duplicate samples. Data are presented asmean±SD. Statistical significance was determined in multiple comparisonsamong independent groups of data in which ANOVA and the F test indicatedthe presence of significant differences. A P value <0.05 was consideredsignificant.

Results The Synergistic Anti-Atherosclerotic Effect of Candesartan andRosuvastatin

Compared with the control mice (C57BL/6J mice fed regular diet), theapo-E knockout mice fed high-cholesterol diet developed extensiveatherosclerosis (P<0.01 vs control mice). Although both candesartan androsuvastatin alone decreased the extent of atherosclerosis (p<0.05 vshigh-cholesterol diet alone), the combination reduced atherosclerosis toa much greater extent (P<0.05 vs candesartan or rosuvastatin alone plushigh-cholesterol diet). FIG. 1 shows results of representativeexperiments and the extent of atherosclerosis (mean±SD) in differentgroups of animals.

Candersartan and rosuvastatin alone decreased atherosclerosis by about35% and 25% respectively. The combination reduced atherosclerosis by70%, demonstrating a synergistic effect. This effect is illustratedgraphically in FIG. 2.

The Synergistic Effect of Candesartan and Rosuvastatin on LOX-1Expression

In the control C57BL/6J mice, the expression of LOX-1 (mRNA and protein)was low. In contrast, LOX-1 expression (mRNA and protein) was markedlyincreased by high-cholesterol diet in apo-E knockout mice (P<0.01 vscontrol mice). Both candesartan and rosuvastatin alone decreased theLOX-1 expression (mRNA and protein), albeit modestly (P<0.05 vshigh-cholesterol diet alone). The combination of candesartan androsuvastatin had a dramatic inhibitory effect on the up-regulation ofLOX-1 (mRNA and protein) in apo-E knockout mice (P<0.01 vshigh-cholesterol diet alone).

The Synergistic Effect of Candesartan and Rosuvastatin on CD40Expression

Compared with the expression in control C57BL/6J mice, CD40 expression(mRNA and protein) was markedly increased in apo-E knockout mice fed ahigh-cholesterol diet in (P<0.01 vs control mice). Although candesartanand rosuvastatin treatment alone slightly decreased CD40 expression(P<0.05 vs high-cholesterol diet alone), the combination of candesartanand rosuvastatin had a dramatic inhibitory effect on the up-regulationof CD40 (mRNA and protein) in the apo-E knockout mice (P<0.01 vshigh-cholesterol diet alone).

The Synergistic Effect of Candesartan and Rosuvastatin on MMPsExpression

Compared with the expression in control C57BL/6J mice, MMP-1, -2 and -9expression (mRNA and protein) was markedly increased in high-cholesteroldiet-fed apo-E knockout mice (P<0.01 vs control mice). Both candesartanand rosuvastatin alone decreased MMP-1, -2 and -9 expression (mRNA andprotein), albeit modestly (P<0.05 vs high-cholesterol diet alone). Thecombination of candesartan and rosuvastatin had a dramatic inhibitoryeffect on their expression (P<0.01 vs high-cholesterol diet alone).

The Effect of Candesartan and Rosuvastatin on TIMPs Expression

TIMP-1 and TIMP-2 protein expression was also increased in apo-Eknockout mice by high-cholesterol diet (P<0.01 vs. control mice), butthe increase was less than that of MMPs. Both candesartan androsuvastatin alone reduced TIMP-1 and TIMP-2 expression by a smalldegree (P<0.05 vs. high-cholesterol diet alone), but the combination ofcandesartan and rosuvastatin had a greater inhibitory effect on theirexpression (P<0.01 vs. high-cholesterol diet alone, P<0.05 vs.high-cholesterol diet with candesartan or rosuvastatin).

1-11. (canceled)
 12. A method of preventing or reducing the risk ofcardiovascular events in a warm-blooded animal in need thereof, whichcomprises administering to said animal an effective amount of acombination comprising candesartan or a pharmaceutically acceptable saltthereof and rosuvastatin or a pharmaceutically acceptable salt thereof.13. The method of claim 12 wherein the combination is administered inassociation with a pharmaceutically acceptable diluent or carrier.
 14. Amethod of preventing or reducing the risk of cardiovascular events in awarm-blooded animal in need thereof, which comprises administering tosaid animal an effective amount of a combination consisting essentiallyof candesartan or a pharmaceutically acceptable salt thereof androsuvastatin or a pharmaceutically acceptable salt thereof as the soletherapeutically effective agents, and optionally a diuretic which ishydrochlorothiazide.
 15. The method of claim 14 wherein the combinationis administered in association with a pharmaceutically acceptablediluent or carrier.
 16. The method of claim 14 wherein the compositionis in the form of a single oral formulation.
 17. The method of any oneof claims 12-15 wherein candesartan is administered in the form ofcandesartan cilexetil.
 18. The method of any one of claims 12-15 whereinthe cardiovascular event is selected from myocardial infarction,worsening of angina, cardiac arrest, stroke, congestive heart failureand cardiovascular death.